Journal
/
Biology
/
Enhanced Crosslinking Immunoprecipitation (eCLIP) Method for Efficient Identification of Protein-bound RNA in Mouse Testis
JoVE Journal
Biology
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biology
Enhanced Crosslinking Immunoprecipitation (eCLIP) Method for Efficient Identification of Protein-bound RNA in Mouse Testis

Enhanced Crosslinking Immunoprecipitation (eCLIP) Method for Efficient Identification of Protein-bound RNA in Mouse Testis

DOI:
10.3791/59681-v

•

10:31 min

•

May 10, 2019

•

, , , , , , , ,
1State Key Laboratory of Reproductive Medicine,Nanjing Medical University

Chapters

  • 00:04Title
  • 01:14Tissue Harvesting and UV Crosslinking
  • 02:37Beads Preparation
  • 03:20Tissue Lysis and Partial RNA Digestion
  • 04:30Immunoprecipitation
  • 05:09RNA Adapter Ligation to RNA 3’ Ends
  • 06:48SDS-PAGE and Membrane Transfer
  • 08:24Results: Analysis of the eCLIP Method
  • 10:08Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we present an eCLIP protocol to determine major RNA targets of RBP candidates in testis.

Tags

Keywords: ECLIPProtein-bound RNAMouse TestisRNA-binding ProteinsSpermatogenesisNon-radioactiveSize-matched InputSmall-scale SequencingCrosslinkingImmunoprecipitationMagnetic Beads

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/59681?article_type=v

Chromatin Immunoprecipitation Assay for Tissue-specific Genes using Early-stage Mouse Embryos
check_url/59681?article_type=v

Efficient Differentiation of Mouse Embryonic Stem Cells into Motor Neurons
check_url/59681?article_type=v

Performing Vaginal Lavage, Crystal Violet Staining, and Vaginal Cytological Evaluation for Mouse Estrous Cycle Staging Identification
check_url/59681?article_type=v

Efficient Chromatin Immunoprecipitation using Limiting Amounts of Biomass
check_url/59681?article_type=v

Identification of Protein Interaction Partners in Mammalian Cells Using SILAC-immunoprecipitation Quantitative Proteomics
check_url/59681?article_type=v

In Vivo Microinjection and Electroporation of Mouse Testis
check_url/59681?article_type=v

Enhanced Northern Blot Detection of Small RNA Species in Drosophila Melanogaster
check_url/59681?article_type=v

Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing
check_url/59681?article_type=v

Chromatin Immunoprecipitation Assay for the Identification of Arabidopsis Protein-DNA Interactions In Vivo
check_url/59681?article_type=v

TChIP-Seq: Cell-Type-Specific Epigenome Profiling

Read Article